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Construct appropriate source of recombinant heavy chain antibodies specific to CD19
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| Author:
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SAMAR KADHUM YOUSEF, AWATIF HAMEED ISSA
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| Abstract:
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Nanobodies (Nbs) are the smallest, functionally integrated unit that has a special importance in the researchers in the field of immunotherapy and biotechnology. These Nbs are derived from heavy chain antibodies only which are found in members of the Camelids, family (llama, camel, alpaca). this paper aimed to establish molecular library of genes encoded to Nbs specific to CD19 ,The immune response was evaluated by ELISA assay ,which revealed a good immune response against the CD-19. Blood collection has performed at 71 days peripheral blood lymphocytes separated by density gradient centrifugation,
cDNA synthesized from immunized lama RNA used as templet for amplify the CD19 -VHH encoding region with the nested polymerase reaction, the first reaction using primers amplification product are 700 bp were detected in 1% agarose gel corresponding to all VH domains of immunoglobulins : IgG2 and IgG3, then fragment at 400 bp was amplified in second PCR reaction,
The second amplification product was cloned into vector pComb3Xss using E.coli Er2738 for transformation confirming the successful of cloning by colony PCR.
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| Keyword:
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CD19, Nanobodies, cloning, Nested PCR, and Lama pacos
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2020.SP2.460
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| Download:
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